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Example sentences for: huape
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The importance of such bonds for activity was shown in other work, where huAPE1's binding to the DNA backbone is only inhibited by ethylation of the phosphates two and three positions 3' to an abasic site [ 18 ] . Mutation of R177A, at the end of Molego 6, that binds to this region and to the bases opposite the AP-site had enhanced activity [ 11 ] , while mutations at W280 (Molego 11) and F266 (Molego 10) [ 33 ] reduce activity and, in the latter case, substrate selectivity.
Mutations that greatly diminish the enzymatic activity of huAPE1 do not affect, and may even increase binding to damaged DNA, while non-specific DNA binding remains low [ 16 18 ] . Further, mutations that have little effect on APE activity in vitro prevent complementation of DNA repair deficient E. coli . As seen with other DNA repair enzymes [ 19 ] , specificity determining residues, as yet unidentified for APEs, must be distinct from those involved in phosphorolysis.
We then demonstrated that the shared molegos served a similar role in substrate binding by comparing the DNA binding profile of huAPE1 with that for the less specific enzyme DNase 1. The molegos present in both enzymes interact with target DNA in a similar fashion, while residues in molegos distinctive for APE1 control specificity by binding primarily to the bases around the apurinic site.
A contact plot of huAPE1 with the DNA in the 1DE8 crystal structure (Figure 5) shows that motifs 1-3,5-8, and 10-12 all have residues close to the substrate, an oligonucleotide containing an abasic site (AP-DNA).
Both enzymes cleave only one DNA strand in a duplex and bacterial Xth cleaves ssDNA containing an abasic site [ 42 ] . The additional contacts huAPE1, compared to DNase 1 (Figure 6), makes 3' to the damage site and to the opposite strand lower its turnover rate and its potential to cleave normal DNA.
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